Hydrogen sulfide improves endothelial barrier function by modulating the ubiquitination degradation of KLF4 through TRAF7 S-sulfhydration in diabetic aorta.

Anomalous vascular endothelium significantly contributes to various cardiovascular diseases. VE-cadherin plays a vital role in governing the endothelial barrier. Krüppel-like factor 4(KLF4), as a transcription factor, which binds the VE-cadherin promoter and enhances its transcription. Tumor necrosis factor receptor-associated factor 7 (TRAF7) is an E3 ubiquitin ligase that has been shown to modulate the degradation of KLF4. H2S can covalently modify cysteine residues on proteins through S-sulfhydration, thereby influencing the structure and functionality of the target protein. However, the role of S-sulfhydration on endothelial barrier integrity remains to be comprehensively elucidated. This study aims to investigate whether protein S-sulfhydration in the endothelium regulates endothelial integrity and its underlying mechanism. In this study, we observed that protein S-sulfhydration was reduced in the endothelium during diabetes and TRAF7 was the main target. Overexpression of TRAF7-Cys327 mutant could mitigate the endothelial barrier damage by weakening TRAF7 interaction with KLF4 and reducing ubiquitination degradation of KLF4. In conclusion, our research demonstrates that H2S plays a pivotal role in regulating S-sulfhydration of TRAF7 at Cys327. This regulation effectively inhibits the ubiquitin-mediated degradation of KLF4, resulting in an upregulation of VE-cadherin levels. This molecular mechanism contributes to the prevention of endothelial barrier damage.

SIRT5-related lysine demalonylation of GSTP1 contributes to cardiomyocyte pyroptosis suppression in diabetic cardiomyopathy.

Sirtuin 5 (SIRT5), localized in the mitochondria, has been identified as a protein desuccinylase and demalonylase in the mitochondria since the depletion of SIRT5 boosted the global succinylation and malonylation of mitochondrial proteins. We investigated the role of SIRT5 in diabetic cardiomyopathy (DCM) and identified the mechanism regarding lysine demalonylation in this process. Wild-type and SIRT5 knockout mice were induced with DCM, and primary cardiomyocytes and cardiac fibroblasts extracted from wild-type and SIRT5 knockout mice were subjected to high glucose (HG). SIRT5 deficiency exacerbated myocardial injury in DCM mice, aggravated HG-induced oxidative stress and mitochondrial dysfunction in cardiomyocytes, and intensified cardiomyocyte senescence, pyroptosis, and DNA damage. DCM-induced SIRT5 loss diminished glutathione S-transferase P (GSTP1) protein stability, represented by significantly increased lysine malonylation (Mal-Lys) modification of GSTP1. SIRT5 overexpression alleviated DCM-related myocardial injury, which was reversed by GSTP1 knockdown. Reduced SIRT5 transcription in DCM resulted from the downregulation of SPI1. SPI1 promoted the transcription of SIRT5, thereby ameliorating DCM-associated myocardial injury. However, SIRT5 deletion resulted in a significant reversal of the protective effect of SPI1. These observations suggest that SPI1 activates SIRT5 transcriptionally to mediate GSTP1 Mal-Lys modification and protein stability, thus ameliorating DCM-associated myocardial injury.

Exogenous H2 S promotes ubiquitin-mediated degradation of SREBP1 to alleviate diabetic cardiomyopathy via SYVN1 S-sulfhydration.

BACKGROUND: Diabetic cardiomyopathy, a distinctive complication of diabetes mellitus, has been correlated with the presence of intracellular lipid deposits. However, the intricate molecular mechanisms governing the aberrant accumulation of lipid droplets within cardiomyocytes remain to be comprehensively elucidated. METHODS: Both obese diabetic (db/db) mice and HL-1 cells treated with 200 µmol/L palmitate and 200 µmol/L oleate were used to simulate type 2 diabetes conditions. Transmission electron microscopy is employed to assess the size and quantity of lipid droplets in the mouse hearts. Transcriptomics analysis was utilized to interrogate mRNA levels. Lipidomics and ubiquitinomics were employed to explore the lipid composition alterations and proteins participating in ubiquitin-mediated degradation in mice. Clinical data were collected from patients with diabetes-associated cardiomyopathy and healthy controls. Western blot analysis was conducted to assess the levels of proteins linked to lipid metabolism, and the biotin-switch assay was employed to quantify protein cysteine S-sulfhydration levels. RESULTS: The administration of H2 S donor, NaHS, effectively restored hydrogen sulfide levels in both the cardiac tissue and plasma of db/db mice (+7%, P < 0.001; +5%, P < 0.001). Both db/db mice (+210%, P < 0.001) and diabetic patients (+83%, P = 0.22, n = 5) exhibit elevated plasma triglyceride levels. Treatment with GYY4137 effectively lowers triglyceride levels in db/db mice (-43%, P = 0.007). The expression of cystathionine gamma-lyase and HMG-CoA reductase degradation protein 1 (SYVN1) was decreased in db/db mice compared with the wild-type mice (cystathionine gamma-lyase: -31%, P = 0.0240; SYVN1: -35%, P = 0.01), and NaHS-treated mice (SYVN1: -31%, P = 0.03). Conversely, the expression of sterol regulatory element-binding protein 1 (SREBP1) was elevated (+91%, P = 0.007; +51%, P = 0.03 compared with control and NaHS-treated mice, respectively), along with diacylglycerol O-acyltransferase 1 (DGAT1) (+95%, P = 0.001; +35%, P = 0.02) and 1-acylglycerol-3-phosphate O-acyltransferase 3 (AGPAT3) (+88%, P = 0.01; +22%, P = 0.32). Exogenous H2 S led to a reduction in lipid droplet formation (-48%, P < 0.001), restoration of SYVN1 expression, modification of SYVN1's S-sulfhydration status and enhancement of SREBP1 ubiquitination. Overexpression of SYVN1 mutated at Cys115 decreased SREBP1 ubiquitination and increased the number of lipid droplets. CONCLUSIONS: Exogenous H2 S enhances ubiquitin-proteasome degradation of SREBP1 and reduces its nuclear translocation by modulating SYVN1's cysteine S-sulfhydration. This pathway limits lipid droplet buildup in cardiac myocytes, ameliorating diabetic cardiomyopathy.

Exogenous H2S initiating Nrf2/GPx4/GSH pathway through promoting Syvn1-Keap1 interaction in diabetic hearts.

Excessive ROS accumulation contributes to cardiac injury in type 2 diabetes mellitus. Hydrogen sulfide (H2S) is a vital endogenous gasotransmitter to alleviate cardiac damage in diabetic cardiomyopathy (DCM). However, the underlying mechanisms remain unclear. In this study, we investigated the effects of NaHS administration in db/db mice via intraperitoneal injection for 20 weeks and the treatment of high glucose (HG), palmitate (PA) and NaHS in HL-1 cardiomyocytes for 48 h, respectively. H2S levels were decreased in hearts of db/db mice and HL-1 cardiomyocytes exposed to HG and PA, which were restored by NaHS. Exogenous H2S activated the nuclear factor erythroid 2-related factor 2 (Nrf2)/glutathione peroxidase 4 (GPx4)/glutathione (GSH) pathway, suppressed ferroptosis and mitigated mitochondrial apoptosis in db/db mice. However, these effects were abrogated after Nrf2 knockdown. NaHS treatment elevated the ubiquitination level of Kelch-like ECH-associated protein (Keap1) by preserving its E3 ligase synoviolin (Syvn1), resulting in Nrf2 nuclear translocation. H2S facilitated the sulfhydration of Syvn1-cys115 site, a post-translational modification. Transfecting Syvn1 C115A in cardiomyocytes exposed to HG and PA partially attenuated the effects of NaHS on Nrf2 and cell death. Our findings suggest that exogenous H2S regulates Nrf2/GPx4/GSH pathway by promoting the Syvn1-Keap1 interaction to reduce ferroptosis and mitochondrial apoptosis in DCM.

Hydrogen sulphide reduced the accumulation of lipid droplets in cardiac tissues of db/db mice via Hrd1 S-sulfhydration.

Accumulation of lipid droplets (LDs) induces cardiac dysfunctions in type 2 diabetes patients. Recent studies have shown that hydrogen sulphide (H2 S) ameliorates cardiac functions in db/db mice, but its regulation on the formation of LDs in cardiac tissues is unclear. Db/db mice were injected with NaHS (40 µmol·kg-1 ) for twelve weeks. H9c2 cells were treated with high glucose (40 mmol/L), oleate (200 µmol/L), palmitate (200 µmol/L) and NaHS (100 µmol/L) for 48 hours. Plasmids for the overexpression of wild-type Hrd1 and Hrd1 mutated at Cys115 were constructed. The interaction between Hrd1 and DGAT1 and DGAT2, the ubiquitylation level of DGAT1 and 2, the S-sulfhydration of Hrd1 were measured. Exogenous H2 S ameliorated the cardiac functions, decreased ER stress and reduced the number of LDs in db/db mice. Exogenous H2 S could elevate the ubiquitination level of DGAT 1 and 2 and increased the expression of Hrd1 in cardiac tissues of db/db mice. The S-sulfhydration of Hrd1 by NaHS enhanced the interaction between Hrd1 and DGAT1 and 2 to inhibit the formation of LD. Our findings suggested that H2 S modified Hrd1 S-sulfhydration at Cys115 to reduce the accumulation of LDs in cardiac tissues of db/db mice.

Exogenous H2 S prevents the nuclear translocation of PDC-E1 and inhibits vascular smooth muscle cell proliferation in the diabetic state.

Hydrogen sulphide (H2 S) inhibits vascular smooth muscle cell (VSMC) proliferation induced by hyperglycaemia and hyperlipidaemia; however, the mechanisms are unclear. Here, we observed lower H2 S levels and higher expression of the proliferation-related proteins PCNA and cyclin D1 in db/db mouse aortae and vascular smooth muscle cells treated with 40 mmol/L glucose and 500 µmol/L palmitate, whereas exogenous H2 S decreased PCNA and cyclin D1 expression. The nuclear translocation of mitochondrial pyruvate dehydrogenase complex-E1 (PDC-E1) was significantly increased in VSMCs treated with high glucose and palmitate, and it increased the level of acetyl-CoA and histone acetylation (H3K9Ac). Exogenous H2 S inhibited PDC-E1 translocation from the mitochondria to the nucleus because PDC-E1 was modified by S-sulfhydration. In addition, PDC-E1 was mutated at Cys101. Overexpression of PDC-E1 mutated at Cys101 increased histone acetylation (H3K9Ac) and VSMC proliferation. Based on these findings, H2 S regulated PDC-E1 S-sulfhydration at Cys101 to prevent its translocation from the mitochondria to the nucleus and to inhibit VSMC proliferation under diabetic conditions.

Numerical Simulation and Experimental Study on Residual Stress in the Curved Surface Forming of 12CrNi2 Alloy Steel by Laser Melting Deposition.

The performance and service life of the nuclear emergency diesel engine shaft made of 12CrNi2 alloy steel is very important for the safety of nuclear power. Laser melting deposition (LMD) is a challenging camshaft-forming technology due to its high precision, rapid prototyping, and excellent parts performance. However, LMD is an unsteady process under the local action of laser, especially for curved surface forming, which is more likely to generate large residual stress on components, resulting in cracks and other defects. At present, the stress research on LMD curved surface forming is relatively insufficient. In the present paper, material parameter testing, high-temperature mechanical properties analysis, single-track sample preparation, and heat source checks are conducted. At the same time, the ABAQUS software and the DFLUX heat source subroutine are used to compile the curved double-ellipsoidal moving heat source, and the effects of the temperature-dependent thermophysical parameters and phase change latent heat on the temperature field are considered. A three-dimensional finite element model is established to analyze the thermal stress evolution and residual stress distribution of multi-track multi-layer on a curved surface by LMD, and the effect of the scanning method and interlayer cooling time on the residual stress of the formed components is studied. The results show that with the increase in temperature, the strength of the material reduces, and the fracture morphology of the material gradually transitions from ductile fracture to creep fracture. The material parameters provide a guarantee for the simulation, and the errors of the width and depth of the melt pool are 4% and 9.6%, respectively. The simulation and experiment fit well. After cooling, the maximum equivalent stress is 686 MPa, which appears at the junction of the substrate and the deposited layer. The larger residual stress is mainly concentrated in the lower part of the deposited layer, where the maximum circumferential stress and axial stress are the tensile stress. Compared with the axial parallel lap scanning method, the arc copying lap scanning method has a relatively smaller maximum thermal stress and residual stress after cooling. The residual stress in the deposited layer is increased to some extent with the increase in the interlayer cooling time.

Hydrogen sulphide ameliorating skeletal muscle atrophy in db/db mice via Muscle RING finger 1 S-sulfhydration.

Muscle atrophy occurs in many pathological states, including cancer, diabetes and sepsis, whose results primarily from accelerated protein degradation and activation of the ubiquitin-proteasome pathway. Expression of Muscle RING finger 1 (MuRF1), an E3 ubiquitin ligase, was increased to induce the loss of muscle mass in diabetic condition. However, hydrogen sulphide (H2 S) plays a crucial role in the variety of physiological functions, including antihypertension, antiproliferation and antioxidant. In this study, db/db mice and C2C12 myoblasts treated by high glucose and palmitate and oleate were chose as animal and cellular models. We explored how exogenous H2 S attenuated the degradation of skeletal muscle via the modification of MuRF1 S-sulfhydration in db/db mice. Our results show cystathionine-r-lyase expression, and H2 S level in skeletal muscle of db/db mice was reduced. Simultaneously, exogenous H2 S could alleviate ROS production and reverse expression of ER stress protein markers. Exogenous H2 S could decrease the ubiquitination level of MYOM1 and MYH4 in db/db mice. In addition, exogenous H2 S reduced the interaction between MuRF1 with MYOM1 and MYH4 via MuRF1 S-sulfhydration. Based on these results, we establish that H2 S prevented the degradation of skeletal muscle via MuRF1 S-sulfhydration at the site of Cys44 in db/db mice.

Inner surface of Nepenthes slippery zone: ratchet effect of lunate cells causes anisotropic superhydrophobicity.

Inner surface of Nepenthes slippery zone shows anisotropic superhydrophobic wettability. Here, we investigate what factors cause the anisotropy via sliding angle measurement, morphology/structure observation and model analysis. Static contact angle of ultrapure-water droplet exhibits the value of 154.80°-156.83°, and sliding angle towards pitcher bottom and up is 2.82 ± 0.45° and 5.22 ± 0.28°, respectively. The slippery zone under investigation is covered by plenty of lunate cells with both ends bending downward, and a dense layer of wax coverings without directional difference in morphology/structure. Results indicate that the slippery zone has a considerable anisotropy in superhydrophobic wettability that is most likely caused by the lunate cells. A model was proposed to quantitatively analyse how the structure characteristics of lunate cells affect the anisotropic superhydrophobicity, and found that the slope/precipice structure of lunate cells forms a ratchet effect to cause ultrapure-water droplet to roll towards pitcher bottom/up in different order of difficulty. Our investigation firstly reveals the mechanism of anisotropic superhydrophobic wettability of Nepenthes slippery zone, and inspires the bionic design of superhydrophobic surfaces with anisotropic properties.

Hydrogen sulfide regulates muscle RING finger-1 protein S-sulfhydration at Cys44 to prevent cardiac structural damage in diabetic cardiomyopathy.

BACKGROUND AND PURPOSE: Hydrogen sulfide (H2 S) plays important roles as a gasotransmitter in pathologies. Increased expression of the E3 ubiquitin ligase, muscle RING finger-1 (MuRF1), may be involved in diabetic cardiomyopathy. Here we have investigated whether and how exogenous H2 S alleviates cardiac muscle degradation through modifications of MuRF1 S-sulfhydration in db/db mice. EXPERIMENTAL APPROACH: Neonatal rat cardiomyocytes were treated with high glucose (40 mM), oleate (100 µM), palmitate (400 µM), and NaHS (100 µM) for 72 hr. MuRF1 was silenced with siRNA technology and mutation at Cys44 . Endoplasmic reticulum stress markers, MuRF1 expression, and ubiquitination level were measured. db/db mice were injected with NaHS (39 µmol·kg-1 ) for 20 weeks. Echocardiography, cardiac ultrastructure, cystathionine-γ-lyase, cardiac structure proteins expression, and S-sulfhydration production were measured. KEY RESULTS: H2 S levels and cystathionine-γ-lyase protein expression in myocardium were decreased in db/db mice. Exogenous H2 S reversed endoplasmic reticulum stress, including impairment of the function of cardiomyocytes and structural damage in db/db mice. Exogenous H2 S could suppress the levels of myosin heavy chain 6 and myosin light chain 2 ubiquitination in cardiac tissues of db/db mice, and MuRF1 was modified by S-sulfhydration, following treatment with exogenous H2 S, to reduce the interaction between MuRF1 and myosin heavy chain 6 and myosin light chain 2. CONCLUSIONS AND IMPLICATIONS: Our findings suggest that H2 S regulates MuRF1 S-sulfhydration at Cys44 to prevent myocardial degradation in the cardiac tissues of db/db mice. LINKED ARTICLES: This article is part of a themed section on Hydrogen Sulfide in Biology & Medicine. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v177.4/issuetoc.

Influence of Crack Size on Stress Evaluation of Ferromagnetic Low Alloy Steel with Metal Magnetic Memory Technology.

Based on the magneto-mechanical effect, the influence of crack size on stress evaluated with metal magnetic memory (MMM) technology was discussed in this paper. Based on equivalent theory, the regular rectangular grooves, with different widths and depths, were precut in the surface of an experimental sample for simulating surface crack, and a three dimensional electrically controlled displacement system was used to collect the Hp(y) signal of the sample under different stresses, and the fracture morphology was observed by using scanning electron microscopy (SEM). The results show that the influence of detection line on Hp(y) signal can be ignored; as stress increases, the Hp(y) signal turns counterclockwise around zero-crossing point and its mutation, corresponding to the location of groove, becomes distinct gradually. When groove depth is constant, the magnetic intensity gradient changes in the form of quadratic polynomial as groove width increases, and when the groove width is the same, the magnetic intensity gradient is a linear function of groove depth. When stress reaches the yield strength of the material, the magnetic intensity gradient decreases gradually as stress increases further, and the orientation of magnetic domain is seen as the main reason for that result. At last, the experimental results are discussed based on the piezomagnetic effect and leakage magnetic field theory of finite depth slit model, and the change of magnetic domain orientation is considered to be the main reason.

Effect of Thermal Cycle on Microstructure Evolution and Mechanical Properties of Selective Laser Melted Low-Alloy Steel.

Low-alloy steel samples were successfully fabricated by selective laser melting (SLM). The evolution of the microstructure and the mechanical properties were investigated with different values of the energy area density (EAD). The results revealed that the initial solidification microstructures of the single tracks with different EADs were all martensite. However, the microstructures of bulk samples under different EADs were not martensite and differed significantly even from one another. When EAD increased from 47 to 142 J/mm2, the mixed lower bainite and martensite austenite microstructure changed to granular bainite; further, the morphology of bainite ferrite gradually changed from lath to multilateral. Moreover, with the increase of EAD, the grain size was remarkably reduced because of the increasing austenitizing periods and temperature during thermal cycling. The average grain size was 1.56 µm, 3.98 µm, and 6.31 µm with EADs of 142 J/mm2, 71 J/mm2, and 47 J/mm2, respectively. Yield strength and tensile strength of the SLM low-alloy steel increased with the increase in EAD; these values were significantly more than those of the alloys prepared by traditional methods. The microstructure of the SLM low-alloy steel samples is not uniform, and the inhomogeneity becomes more significant as EAD decreases. Simultaneously, when EAD decreases, the fracture mechanism changes from ductile to a mixture of ductile and brittle fracture; this is in contrast to the samples prepared by traditional methods. This study also found a stress concentration mechanism around large pores during plastic deformation that resulted in a brittle fracture. This indicates that large-sized pores significantly degrade the mechanical properties of the specimens.

CaSR activates PKCδ to induce cardiomyocyte apoptosis via ER stress­associated apoptotic pathways during ischemia/reperfusion.

Endoplasmic reticulum (ER) stress can be activated by ischemia/reperfusion (I/R) injury in cardiomyocytes. Persistent ER stress, with an increase in intracellular Ca2+ ([Ca2+]i) concentration, leads to apoptosis. Protein kinase C (PKC) has a key role in myocardial damage by elevation of [Ca2+]i. The calcium­sensing receptor (CaSR), a G protein­coupled receptor, can increase the release of [Ca2+]i from the ER through the inositol triphosphate receptor (IP3R). Intracellular calcium overload has been demonstrated to cause cardiac myocyte apoptosis during I/R. However, the associations between PKC, CaSR and ER stress are not clear. The present study examined the hypothesis that activation of PKCδ by CaSR participates in ER stress­associated apoptotic pathways within myocardial I/R. Rat hearts were subjected to 30 min of ischemia in vivo, followed by reperfusion for 120 min. GdCl3 (a CaSR activator) was used to elevate the intracellular Ca2+ concentration, but the Ca2+ concentration in the ER was significantly decreased during I/R. Following exposure to GdCl3, expression levels of CaSR, glucose­regulated protein 78 (GRP78), Caspase­12, phosphorylated JNK and Caspase­3 were increased, and the ratios of apoptotic myocardial cells were significantly increased. By contrast, following exposure to rottlerin, a PKCδ inhibitor, the expression levels of these proteins and the ratio of apoptotic myocardial cells were significantly reduced. The present study also demonstrated that PKCδ translocated into the ER to induce an ER stress response and participate in the ER stress­related apoptosis pathway. These results confirmed that CaSR activated PKCδ to induce cardiomyocyte apoptosis through ER stress­associated apoptotic pathways during I/R in vivo.

Exogenous H2S reduces the acetylation levels of mitochondrial respiratory enzymes via regulating the NAD+-SIRT3 pathway in cardiac tissues of db/db mice.

Hydrogen sulfide (H2S), a gaseous molecule, is involved in modulating multiple physiological functions, such as antioxidant, antihypertension, and the production of polysulfide cysteine. H2S may inhibit reactive oxygen species generation and ATP production through modulating respiratory chain enzyme activities; however, the mechanism of this effect remains unclear. In this study, db/db mice, neonatal rat cardiomyocytes, and H9c2 cells treated with high glucose, oleate, and palmitate were used as animal and cellular models of type 2 diabetes. The mitochondrial respiratory rate, respiratory chain complex activities, and ATP production were decreased in db/db mice compared with those in db/db mice treated with exogenous H2S. Liquid chromatography with tandem mass spectrometry analysis showed that the acetylation level of proteins involved in the mitochondrial respiratory chain were increased in the db/db mice hearts compared with those with sodium hydrosulfide (NaHS) treatment. Exogenous H2S restored the ratio of NAD+/NADH, enhanced the expression and activity of sirtuin 3 (SIRT3) and decreased mitochondrial acetylation level in cardiomyocytes under hyperglycemia and hyperlipidemia. As a result of SIRT3 activation, acetylation of the respiratory complexe enzymes NADH dehydrogenase 1 (ND1), ubiquinol cytochrome c reductase core protein 1, and ATP synthase mitochondrial F1 complex assembly factor 1 was reduced, which enhanced the activities of the mitochondrial respiratory chain activity and ATP production. We conclude that exogenous H2S plays a critical role in improving cardiac mitochondrial function in diabetes by upregulating SIRT3.

Antifouling performance and mechanism of elastic graphene-silicone rubber composite membranes.

Composite coatings have attracted great attention as an eco-friendly and economic solution to prevent ship hulls from biofouling. Inspired by the unstable surfaces of marine organisms with antifouling properties, this study describes the preparation of graphene-silicone rubber composite membranes. The membranes are characterized by a low surface energy and an adjustable elastic modulus, and these properties are conducive to preventing biofouling. Bacterial attachment was tested under both quasi-static and hydrodynamic conditions, and one rigid polystyrene sheet was used as the control group to verify the antifouling effects of unstable surfaces. The polystyrene sheet and the elastic membranes showed similar antifouling performance under quasi-static conditions. However, under hydrodynamic conditions, the elastic membranes showed better antifouling performance than the rigid polystyrene sheet. The results obtained using a laser-displacement sensor showed that micron-scale deformations were present on the elastic surface, and a mechanical model was employed to verify this conclusion. This study first confirmed the antifouling effects of the unstable surface, and proposed a model to reveal the antifouling mechanism of the unstable surface. According to the bacterial attachment test, a new generation membrane was made showing antifouling capacity with just 0.36 wt% graphene included during the fabrication of the membrane. This study provided a deeper insight into the antifouling mechanism of the elastic surface, and the membrane (0.36 wt%) may be promising for practical applications.

Progress and Challenges of Ultrasonic Testing for Stress in Remanufacturing Laser Cladding Coating.

Stress in laser cladding coating is an important factor affecting the safe operation of remanufacturing components. Ultrasonic testing has become a popular approach in the nondestructive evaluation of stress, because it has the advantages of safety, nondestructiveness, and online detection. This paper provides a review of ultrasonic testing for stress in remanufacturing laser cladding coating. It summarizes the recent research outcomes on ultrasonic testing for stress, and analyzes the mechanism of ultrasonic testing for stress. Remanufacturing laser cladding coating shows typical anisotropic behaviors. The ultrasonic testing signal in laser cladding coating is influenced by many complex factors, such as microstructure, defect, temperature, and surface roughness, among others. At present, ultrasonic testing for stress in laser cladding coating can only be done roughly. This paper discusses the active mechanism of micro/macro factors in the reliability of stress measurement, as well as the impact of stress measurement on the quality and safety of remanufacturing components. Based on the discussion, this paper proposes strategies to nondestructively, rapidly, and accurately measure stress in remanufacturing laser cladding coating.

Exogenous H2S switches cardiac energy substrate metabolism by regulating SIRT3 expression in db/db mice.

Hydrogen sulfide (H2S) is involved in diverse physiological functions, such as anti-hypertension, anti-proliferation, regulating ATP synthesis, and reactive oxygen species production. Sirtuin 3 (SIRT3) is a NAD + -dependent deacetylase that regulates mitochondrial energy metabolism. The role of H2S in energy metabolism in diabetic cardiomyopathy (DCM) may be related to regulate SIRT3 expression; however, this role remains to be elucidated. We hypothesized that exogenous H2S could switch cardiac energy metabolic substrate preference by lysine acetylation through promoting the expression of SIRT3 in cardiac tissue of db/db mice. Db/db mice, neonatal rat cardiomyocytes, and H9c2 cell line with the treatment of high glucose, oleate, and palmitate were used as animal and cellular models of type 2 diabetes. Using LC-MS/MS, we identified 76 proteins that increased acetylation, including 8 enzymes related to fatty acid ß-oxidation and 7 enzymes of the tricarboxylic acid (TCA) cycle in the db/db mice hearts compared to those with the treatment of NaHS. Exogenous H2S restored the expression of NAMPT and the ratio of NAD+/NADH enhanced the expression and activity of SIRT3. As a result of activation of SIRT3, the acetylation level and activity of fatty acid ß-oxidation enzyme LCAD and the acetylation of glucose oxidation enzymes PDH, IDH2, and CS were reduced which resulted in activation of PDH, IDH2, and CS. Our finding suggested that H2S induced a switch in cardiac energy substrate utilization from fatty acid ß-oxidation to glucose oxidation in DCM through regulating SIRT3 pathway. KEY MESSAGES: H2S regulated the acetylation level and activities of enzymes in fatty acid oxidation and glucose oxidation in cardiac tissues of db/db mice. Exogenous H2S decreased mitochondrial acetylation level through upregulating the expression and activity of SIRT3 in vivo and in vitro. H2S induced a switch in cardiac energy substrate utilization from fatty acid oxidation to glucose.

Contributions of lunate cells and wax crystals to the surface anisotropy of Nepenthes slippery zone.

Nepenthes slippery zone presents surface anisotropy depending on its specialized structures. Herein, via macro-micro-nano scaled experiments, we analysed the contributions of lunate cells and wax crystals to this anisotropy. Macroscopic climbing of insects showed large displacements (triple body length within 3 s) and high velocities (6.16-20.47 mm s-1) in the inverted-fixed (towards digestive zone) slippery zone, but failed to climb forward in the normal-fixed (towards peristome) one. Friction force of insect claws sliding across inverted-fixed lunate cells was about 2.4 times of that sliding across the normal-fixed ones, whereas showed unobvious differences (1.06-1.11 times) between the inverted- and normal-fixed wax crystals. Innovative results from atomic force microscope scanning and microstructure examination demonstrated the upper layer of wax crystals causes the cantilever tip to generate rather small differences in friction data (1.92-2.72%), and the beneath layer provides slightly higher differences (4.96-7.91%). The study confirms the anisotropic configuration of lunate cells produces most of the anisotropy, whereas both surface topography and structural features of the wax crystals generate a slight contribution. These results are helpful for understanding the surface anisotropy of Nepenthes slippery zone, and guide the design of bioinspired surface with anisotropic properties.

Exogenous H2S Protects Against Diabetic Cardiomyopathy by Activating Autophagy via the AMPK/mTOR Pathway.

BACKGROUND/AIM: Autophagy plays an important role in cellular homeostasis through the disposal and recycling of cellular components. Hydrogen sulphide (H2S) is the third endogenous gas that has been shown to confer cardiac protective effects. Given the regulation of autophagy in cardioprotection, this study aimed to investigate the protective effects of H2S via autophagy during high glucose treatment. METHODS: This study investigated the content of H2S in the plasma as well as myocardial, ultrastructural changes in mitochondria and autophagosomes. This study also investigated the apoptotic rate using Hoechst/PI as well as expression of autophagy-associated proteins and mitochondrial apoptotic proteins in H9C2 cells treated with or without GYY4137. Mitochondria of cardiac tissues were isolated and RCR and ADP/O were also detected. AMPK knockdown was performed with siRNA transfection. RESULTS: In a STZ-induced diabetic model, NaHS treatment not only increased the expression of p-AMPK in diabetic group but further activated cell autophagy. Following 48h high glucose, autophagosomes and cell viability were reduced. The present results showed that autophagy could be induced by H2S, which was verified by autophagic ultrastructural observation and LC3-I/LC3-II conversion. In addition, the mitochondrial membrane potential (MMP) was significantly decreased. The expressions levels of autophagic-related proteins were significantly elevated. Moreover, H2S activated the AMPK/rapamycin (mTOR) signalling pathway. CONCLUSIONS: Our findings demonstrated that H2S decreases oxidative stress and protects against mitochondria injury, activates autophagy, and eventually leads to cardiac protection via the AMPK/mTOR pathway.

Exogenous H2S facilitating ubiquitin aggregates clearance via autophagy attenuates type 2 diabetes-induced cardiomyopathy.

Diabetic cardiomyopathy (DCM) is a serious complication of diabetes. Hydrogen sulphide (H2S), a newly found gaseous signalling molecule, has an important role in many regulatory functions. The purpose of this study is to investigate the effects of exogenous H2S on autophagy and its possible mechanism in DCM induced by type II diabetes (T2DCM). In this study, we found that sodium hydrosulphide (NaHS) attenuated the augment in left ventricular (LV) mass and increased LV volume, decreased reactive oxygen species (ROS) production and ameliorated H2S production in the hearts of db/db mice. NaHS facilitated autophagosome content degradation, reduced the expression of P62 (a known substrate of autophagy) and increased the expression of microtubule-associated protein 1 light chain 3 II. It also increased the expression of autophagy-related protein 7 (ATG7) and Beclin1 in db/db mouse hearts. NaHS increased the expression of Kelch-like ECH-associated protein 1 (Keap-1) and reduced the ubiquitylation level in the hearts of db/db mice. 1,4-Dithiothreitol, an inhibitor of disulphide bonds, increased the ubiquitylation level of Keap-1, suppressed the expression of Keap-1 and abolished the effects of NaHS on ubiquitin aggregate clearance and ROS production in H9C2 cells treated with high glucose and palmitate. Overall, we concluded that exogenous H2S promoted ubiquitin aggregate clearance via autophagy, which might exert its antioxidative effect in db/db mouse myocardia. Moreover, exogenous H2S increased Keap-1 expression by suppressing its ubiquitylation, which might have an important role in ubiquitin aggregate clearance via autophagy. Our findings provide new insight into the mechanisms responsible for the antioxidative effects of H2S in the context of T2DCM.